This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Net1 sequesters the Cdc14 protein phosphatase and other proteins in the yeast nucleolus through much of the cell cycle. The Cdc5 and Cdc28 protein kinases regulate release of Cdc14 from the Net1 complex. We would like to use mass spectrometry to identify other proteins that are released from the Net1 complex upon phosphorylation by additional kinases. This study should provide invaluable information on how multiple kinases regulate the sequestration of proteins in the nucleolus.